中草药中胰高血糖素受体拮抗剂的高通量细胞筛选  

High Throughput Screening Method of Identifying Antagonist for Glucagaon Receptor in Chinese Herbs

作　　者：万娟[1] 许治良[1] 

机构地区：[1]华东师范大学生命科学学院上海市脑功能基因组学重点实验室,上海200062

出　　处：《华东师范大学学报（自然科学版）》2007年第4期112-118,共7页Journal of East China Normal University(Natural Science)

基　　金：国家科技部863计划项目(2003AA221061)

摘　　要：为筛选胰高血糖素受体拮抗剂,建立了一种高通量筛选胰高血糖素受体拮抗剂的细胞模型.利用PCR方法扩增得到人的胰高血糖素受体cDNA,并连接到哺乳动物细胞表达载体pCDNA3.1上,构成质粒h glucagons-R/pCDNA3.1,将这种质粒与报告基因质粒(3×CRE/3×MRE/SRE-LUC)共转染到HEK293细胞,通过G418筛选,建立稳定的人胰高血糖素受体拮抗剂筛选细胞株.利用胰高血糖素受体内源激动剂探索和优化了筛选条件,筛选方法合理,Z′因子大于0.6,系统稳定,符合胰高血糖素受体拮抗剂的高通量筛选的要求.This paper generated a cell-based functional assay for human glucagon receptor antagonist screening. The human glucagon receptor cDNA was cloned into a mammalian expression vector pCDNA3.1 （h Glucagon receptor/pCDNA3. 1）. This plasmid was co transfected with a reporter gene construct （3×CRE/3× MRE/SRE-LUC） in HEK293 cells. Stable cell line expressing both human glucagon receptor and the reporter gene was selected for compounds screening. The assay condition was optimized. Our results demonstrated that this cell-based assay can be optimized for high throughput screening and applied to identify antagonist for glucagon receptor in Chinese Herbs.

关 键 词：胰高血糖素受体 报告基因 拮抗剂 高通量药物筛选 

分 类 号：R965[医药卫生—药理学]